This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Recent findings suggest that local protein synthesis is required for axon navigation in the brain. Axon guidance molecules such as netrin-1, semaphorin 3A and BDNF, elicit chemotropic turning responses in retinal growth cones. These guidance molecules trigger rapid protein synthesis (within 10 min.) in retinal growth cones and, importantly, inhibition of protein synthesis blocks directional responses (Campbell and Holt, 2001). Recent preliminary data indicate that ?-actin is rapidly synthesised in response to guidance cues raising the possibility that rapid local synthesis of cytoskeletal proteins is triggered by guidance cues and is necessary for turning. The goal of the present collaborative project is to investigate the role of the local synthesis of cytoskeletal proteins (?-actin and tubulin) in growth cone steering. We plan to use the FLAsh-ReAsh system to visualize the dynamic synthesis of ?-actin and tubulin in growth cones in response to guidance cues. In particular, we shall address whether a gradient of netrin-1 causes assymmetric synthesis of ?-actin and or tubulin in the growth cone.
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